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J. Biol. Chem., Vol. 278, Issue 34, 32204-32211, August 22, 2003

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The NOG1 GTP-binding Protein Is Required for Biogenesis of the 60 S Ribosomal Subunit^*

Bryan C. Jensen   ¶, Qin Wang  , Charles T. Kifer  and Marilyn Parsons   ||

From the Seattle Biomedical Research Institute, Seattle, Washington 98109 and Department of Pathobiology, School of Community Medicine and Public Health, University of Washington, Seattle, Washington 98195

NOG1 is a nucleolar GTP-binding protein present in eukaryotes ranging from trypanosomes to humans. In this report we demonstrate that NOG1 is functionally linked to ribosome biogenesis. In sucrose density gradients Trypanosoma brucei NOG1 co-sediments with 60 S ribosomal subunits but not with monosomes. 60 S precursor RNAs are co-precipitated with NOG1. Together with the nucleolar localization of NOG1, these data indicate that NOG1 is associated with a precursor particle to the 60 S subunit. Disruption of NOG1 function through RNA interference led to a dramatic decrease in the levels of free 60 S particles and the appearance of an atypical rRNA intermediate in which ITS2 was not cleaved. Overexpression of mutant nog1 with a defect in its GTP binding motif on a wild type background caused a modest defect in 60 S biogenesis and a relative decrease in processing of the large subunit rRNAs. In contrast to the mutant protein, neither the N-terminal half of NOG1, which contains the GTP binding motifs, nor the C-terminal half of NOG1 associated with pre-ribosomal particles, although both localized to the nucleolus.

Received for publication, April 22, 2003

Note Added in Proof—During review of this report, NOGI was shown to be required for 60 S biogenesis in yeast (48).

^* This work was supported in part by National Institutes of Heath Grant R01 AI31077. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Supported by National Institutes of Heath Grant 1T32 AI07509.

^|| To whom correspondence should be addressed: Seattle Biomedical Research Institute, 4 Nickerson St., Seattle, WA 98109. Tel.: 206-284-8846 (ext. 315); Fax: 206-284-0313; E-mail: mparsons{at}sbri.org.

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